>We wish to transfect mRNA (synthesized in vitro) into a transformed rat cell
>line and are seeking advice and suggestions from those who've had experience
>doing this.
>>Thanks in advance,
>>Anthony F.
We transfect in vitro transcriptions via electroporation, in RNAse-free PBS.
The conditions will have to be worked out on your machine with your cells.
We have calculated that ~60% of survivors (BHK-21 cells) take up the RNA.
Also, lipofectin works well for RNA, albeit at lower efficiency than
electroporation. Again, you'll have to optimize conditions. Note, lipofectamine
was far less efficient than lipofectin for delivering RNA, although the reverse
has generally been true for DNA in our lab. Just my thoughts...
Brett Lindenbach
Program in Immunology
Washington University - St Louis
brett at borcim.wustl.edu
"I own my own pet virus. I get to pet and name her." - Cobain